| Title |
Recombinant expression and purification of the 2,5-diketocamphane 1,2-monooxygenase from the camphor metabolizing Pseudomonas putida strain NCIMB 10007
|
|---|---|
| Published in |
AMB Express, June 2011
|
| DOI | 10.1186/2191-0855-1-13 |
| Pubmed ID | |
| Authors |
Maria Kadow, Stefan Saß, Marlen Schmidt, Uwe T Bornscheuer |
| Abstract |
Three different Baeyer-Villiger monooxygenases (BVMOs) were reported to be involved in the camphor metabolism by Pseudomonas putida NCIMB 10007. During (+)-camphor degradation, 2,5-diketocamphane is formed serving as substrate for the 2,5-diketocamphane 1,2-monooxygenase. This enzyme is encoded on the CAM plasmid and depends on the cofactors FMN and NADH and hence belongs to the group of type II BVMOs. We have cloned and recombinantly expressed the oxygenating subunit of the 2,5-diketocamphane 1,2-monooxygenase (2,5-DKCMO) in E. coli followed by His-tag-based affinity purification. A range of compounds representing different BVMO substrate classes were then investigated, but only bicyclic ketones were converted by 2,5-DKCMO used as crude cell extract or after purification. Interestingly, also (-)-camphor was oxidized, but conversion was about 3-fold lower compared to (+)-camphor. Moreover, activity of purified 2,5-DKCMO was observed in the absence of an NADH-dehydrogenase subunit. |
Mendeley readers
The data shown below were compiled from readership statistics for 22 Mendeley readers of this research output. Click here to see the associated Mendeley record.
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 22 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Researcher | 4 | 18% |
| Student > Ph. D. Student | 4 | 18% |
| Student > Bachelor | 3 | 14% |
| Student > Master | 3 | 14% |
| Student > Doctoral Student | 1 | 5% |
| Other | 1 | 5% |
| Unknown | 6 | 27% |
| Readers by discipline | Count | As % |
|---|---|---|
| Agricultural and Biological Sciences | 7 | 32% |
| Biochemistry, Genetics and Molecular Biology | 6 | 27% |
| Immunology and Microbiology | 1 | 5% |
| Materials Science | 1 | 5% |
| Engineering | 1 | 5% |
| Other | 0 | 0% |
| Unknown | 6 | 27% |
Attention Score in Context
This research output has an Altmetric Attention Score of 3. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 03 February 2013.
All research outputs
#7,454,951
of 22,790,780 outputs
Outputs from AMB Express
#190
of 1,234 outputs
Outputs of similar age
#41,552
of 115,205 outputs
Outputs of similar age from AMB Express
#2
of 6 outputs
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So far Altmetric has tracked 1,234 research outputs from this source. They receive a mean Attention Score of 2.8. This one has done well, scoring higher than 82% of its peers.
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